1st Edition
Practical Fluorescence Spectroscopy
Presenting a detailed, hands-on approach to fluorescence spectroscopy, this book describes experiments that cover basic spectroscopy and advanced aspects of fluorescence spectroscopy. It emphasizes practical guidance, providing background on fundamental concepts as well as guidance on how to handle artifacts, avoid common errors, and interpret data. Nearly 150 experiments from biophysics, biochemistry, and the biomedical sciences demonstrate how methods are applied in practical applications. The result is a hands-on guide to the most important aspects of fluorescence spectroscopy, from steady-state fluorescence to advanced time-resolved fluorescence.
- Provides a complete overview of nearly 150 experiments using fluorescence spectroscopy, from basic to advanced applications
- Presents laboratory methods using a variety of instrumental setups with detailed discussion of data analysis and interpretations
- Covers steady-state phenomena, time-resolved phenomena, and advanced methods
- Spans biophysical, biochemical, and biomedical applications
- Describes related concepts, theory, and mathematical background as well as commercially available instruments used for measurements
Chapter 1. Theory of Light and Light Interaction with Matter
Basics of Light
Light Polarization
Interaction of Light and Chromophores
Interactions Among Chromphores
Scattering
Summary
Chapter 2. Experimental Basics
Types of Cuvettes
Types of Filters
Measuring Transmittance/Absorbance
Factors Affecting the Precision of Absorption Measurement
Effect of Sample Scattering on Measured Transmission and Abosrbance
Measuring Emission
Efficiency of Collecting Emission: Measuring Quantum Yield
Wavelength Calibration
Instrucment Response
Lamp Profile
Peaks and Spectral Shapes
Standard Deviation and Height
Sums of Peaks
Noise–Signal-to-Noise Ratio
Sampling Rate
Bandwidth
Artifacts and Errors
Spectral Overlap
Peak Normalization
Residuals
Interpolation
Chapter 3. Experiments
Introduction
Experiments
Chapter 4. Fluorescence–Steady-State Phenomena
Introduction
Experiments
Chapter 5. Steady-State Fluorescence: Applications
Introduction
Experiments
Chapter 6. Steady-State Fluorescence Polarization: Anisotropy
Introduction
How to Select and Check Polarizers
Transition Moments
Polarized Absorption
Emission Polarization (Anisotropy)
Fundamental and Limiting Anistropies
Consequences of Linear Transition Moments
Determining the G-Factor for Spectrofluorometer
Experiments
Chapter 7. Fluorescence: Time-Resolved Phenomena
Introduction
Experiments
Chapter 8. Advanced Experiments
Introduction
Experiments
Biography
Zygmunt (Karol) Gryczynski is a professor of molecular biology and immunology and the director of the Center for Commercialization of Fluorescence Technologies (CCFT) the University of North Texas Health Center.
Ignacy Gryczynski is a professor of cell biology and genetics and the director of the Microscopy Core Facility at the University of North Texas Health Center.
